Deep topical systemic nitric oxide therapy apparatus and method

ABSTRACT

A topical mixture that produces nitric oxide and a method for using the topical mixture to increase the vasodilation of a bloodstream via transdermal absorption of the nitric oxide. The nitric oxide can then affect subcutaneous tissues. The systemic vasodilation of a mammal may be increased via a topical application of an appropriate nitric oxide producing substance.

RELATED APPLICATIONS

This patent application is a continuation of U.S. patent applicationSer. No. 17/082,709, filed Oct. 28, 2020, which is a divisional of U.S.patent application Ser. No. 15/492,528, filed Apr. 20, 2017, issued asU.S. Pat. No. 10,828,249 on Nov. 10, 2020; which is a divisional of U.S.patent application Ser. No. 14/274,419, filed May 9, 2014, issued asU.S. Pat. No. 10,517,817 on Dec. 31, 2019; which claims the benefit ofU.S. Provisional Patent Application Ser. No. 61/821,443, filed May 9,2013; U.S. Provisional Patent Application Ser. No. 61/888,415, filedOct. 8, 2013; and U.S. Provisional Patent Application Ser. No.61/990,591, filed May 8, 2014, which are hereby incorporated byreference in their entireties.

BACKGROUND Field of the Invention

This invention relates to generation of nitric oxide (NO), and moreparticularly, topical application treatments providing nitric oxide as avasodilator, including transdermal penetration of nitric oxide to deeptissues and blood vessels resulting in vasodilation.

Description of the Related Art

The discovery of the nitric oxide effect in live tissues garnered aNobel prize. Much of the work in determining the mechanisms forimplementing and the effects of nitric oxide administration are reportedin literature including papers, advertising, catalogs, and patents.

Much of the work deals with introduction of substances that provide anitric oxide effect in the body. Other applications involve productionand use of nitric oxide to sterilize a variety of surfaces, including aperson's skin. As an example, U.S. Pat. No. 8,501,090 disclosesproduction and use of nitric oxide for sterilization purposes, whichpatent is hereby incorporated by reference in its entirety. Still otherapplications may involve topical preparations introducing nitric oxide.Still other applications rely on bottled nitric oxide gas. Introductionof nitric oxide to the human body has traditionally been expensive andsometimes difficult to achieve or substantiate.

The therapies, compositions, and preparations for promoting nitric oxideproduction in the body can be sufficiently expensive to inhibit morewidespread use of such therapies, and can lose effectiveness. Forexample, L-arginine supplements can increase the main precursor forendothelial nitric oxide synthase to produce nitric oxide, which canimprove vascular reactivity and functional capacity in the short term,but in the long term can result in no increase in endothelial nitricoxide synthase or nitric oxide, which can decrease vascular reactivityand functional capacity. Nitric oxide donors (systemicnitrovasodilators) can help preserve endothelial function and increasenitric oxide, but a tolerance can be developed, including generatingoxidative stress causing further endothelial dysfunction.

What is needed is a comparatively inexpensive mechanism for introducingnitric oxide in a single dosage over an extended period of time. Also,what is needed is a simple, topical application method to promotetransdermal absorption of nitric oxide subcutaneously, into deepertissues of the body, and increase vasodilation. Also, what is needed isa simple, topical application method to promote transdermal absorptionof nitric oxide in order to gain numerous other benefits from suchnitric oxide absorption in subcutaneous tissues.

Topical uses of nitric oxide producing gels and mediums have includedsurface uses and applications of such gels and mediums, includinganti-septic, sterilization, and wound healing. What is needed is atopical use directed toward transdermal, sub-cutaneous absorption ofnitric oxide through healthy skin into the deeper, subcutaneous tissuesof the body to promote vasodilation, as well as other benefits fromnitric oxide, including but not limited to, anti-inflammatory,antiseptic, microbicide, biofilm dispersal, and analgesic.

SUMMARY OF THE INVENTION

In accordance with the foregoing, certain embodiments of an apparatusand method in accordance with the invention provide a system forgenerating nitric oxide (NO) having two compounds, typically disposed inseparate carriers. The two compounds are separated from one anotherprior to administration. In order to administer the nitric oxide,reactants are mixed with one another, beginning a reaction releasingnitric oxide. The resultant mixture is applied to the skin so the nitricoxide can be absorbed transdermally into the bloodstream and promotevasodilation of subcutaneous tissues.

In certain embodiments, a topical mixture may include a first medium anda second medium. The first medium may include a nitrite salt capable ofproviding nitric oxide upon reaction with an acid or reducing agent, forexample, the nitrite salt may be sodium nitrite, potassium nitrite, orthe like. The second medium may include an acid or reducing agent forreacting with the nitrite salt, for example, the acid or reducing agentmay be citric acid, ascorbic acid, phytic acid, acetic acid, formicacid, lactic acid, or the like. The mixture of the first medium and thesecond medium may initiate production of nitric oxide.

The mixture may also include enough reactants to produce an effectiveamount of nitric oxide sufficient to increase vasodilation, systemicvasodilation, and nitric oxide levels in a mammal's bloodstream viatransdermal, subcutaneous absorption.

In certain embodiments, a method for increasing nitric oxide levels inthe bloodstream via transdermal, subcutaneous absorption may includeproviding a mammal with an epidermal or skin surface, wherein the mammalalso has an initial amount of nitric oxide in its bloodstream and aninitial systemic vasodilation of the vascular system. Then, a topicalmixture that may include a first medium and a second medium is appliedto the skin of the mammal. The first medium may include a nitrite saltcapable of providing nitric oxide upon reaction with an acid or reducingagent, for example, the nitrite salt may be sodium nitrite, potassiumnitrite, or the like. The second medium may include an acid or reducingagent for reacting with the nitrite salt, for example, the acid orreducing agent may be citric acid, ascorbic acid, phytic acid, aceticacid, formic acid, lactic acid, or the like. The mixture of the firstmedium and the second medium may initiate production of nitric oxide.Thus, the amount of nitric oxide in the bloodstream of the mammal isincreased and the systemic vasodilation of its vascular system isincreased.

In certain embodiments, the epidermal surface or skin of the mammal maybe healthy skin. Also, the epidermal surface of the skin of the mammalmay include acne. Also, the epidermal surface of the skin of the mammalmay include a biofilm. Also, the skin may be covered with a barrier,cover, or bandage after application of the nitric oxide productionmixture.

In certain embodiments, the mixture of reactants may include a bufferconcentration that is included to prolong the production of nitricoxide, for example, a salt buffer of the like. A salt like sodiumchloride, calcium chloride, or the like, may be added to the firstmedium, second medium, or both in a sufficient amount to inhibit or slowthe reaction between the nitrite salt and the reducing agent.

Also, the mixture of reactants may include thickening agents to increasethe viscosity of the mixture, or the viscosity of one or both of theunderlying, initial mediums, to prolong the production of nitric oxide,for example, hydroxyethyl cellulose, methyl cellulose, hydroxypropylmethylcellulose, or the like.

In accordance with the present invention, many ailments or diseases maybe treated by transdermal, topical mixture applications producing nitricoxide, including without limitation, peripheral artery disease,hypertension, diabetes, atherosclerosis, acne, inflammation, tendonitis,biofilms, and the like.

In certain embodiments, a method for increasing vasodilation in a mammalvia transdermal, sub-cutaneous absorption of nitric oxide may includeproviding a mammal with an epidermal or skin surface, wherein the mammalalso has an initial level of vasodilation, or systemic vasodilation, inits vascular system. Then, a topical mixture that may include a firstmedium and a second medium is applied to the skin of the mammal. Thefirst medium may include a nitrite salt capable of providing nitricoxide upon reaction with an acid or reducing agent, for example, thenitrite salt may be sodium nitrite, potassium nitrite, or the like. Thesecond medium may include an acid or reducing agent for reacting withthe nitrite salt, for example, the acid or reducing agent may be citricacid, ascorbic acid, phytic acid, acetic acid, formic acid, lactic acid,or the like. The mixture of the first medium and the second medium mayinitiate production of nitric oxide. Thus, the systemic vasodilation ofthe mammal is increased after transdermal absorption of nitric oxide.

Certain embodiments of an apparatus and method in accordance with theinvention provide a topical medium that produces nitric oxide andprovides a therapeutic concentration of nitric oxide delivered to asurface. Nitric oxide may thus be introduced to the skin, or a wound, ofa subject in a controlled manner. Nitric oxide amounts may be engineeredto deliver a therapeutically effective amount on the order of fromcomparatively low hundreds (e.g., 100-500) of parts per million, up tothousands of parts per million (ppm). For example, sufficient nitricoxide may be presented through topical application to provideapproximately five hundred parts per million to the surface of asubject's skin, and up to 2500 ppm.

One embodiment of an apparatus and method in accordance with the presentinvention may rely on equal amounts of a nitrite medium and an acidifiedmedium formulated to provide a burst of nitric oxide, as well as acontinuous amount of nitric oxide over a period of time.

One embodiment of an apparatus and method in accordance with the presentinvention may provide a therapeutically effective amount of nitric oxidefrom a gel medium, which provides a therapeutically effective dose ofnitric oxide over a potentially shorter length of time, fromapproximately ten to thirty minutes, and potentially up to about 3hours. The reaction may begin upon combination of the nitrite medium andan acidified gel medium.

One embodiment of an apparatus and method in accordance with the presentinvention may provide a therapeutically effective amount of nitric oxidefrom a lotion medium, which provides a therapeutically effective dose ofnitric oxide over a potentially longer length of time, from about onehour up to about 6 hours. Reactants may include potassium nitrite,sodium nitrite or the like. The reaction may begin upon combination ofthe nitrite medium and an acidified medium.

One embodiment of an apparatus and method in accordance with the presentinvention may provide a therapeutically effective amount of nitric oxidefrom a serum medium, which provides a therapeutically effective dose ofnitric oxide over limited time, from about initial application to thirtyminutes or sixty minutes. Reactants may include potassium nitrite,sodium nitrite or the like. The reaction may begin upon combination ofthe nitrite serum medium and the acidified serum medium.

One embodiment of an apparatus and method in accordance with the presentinvention may provide a therapeutically effective amount of nitric oxidefrom a mud (facial mud) medium, which provides a therapeuticallyeffective dose of nitric oxide over limited time, from about initialapplication from about one hour to about three hours. Reactants mayinclude potassium nitrite, sodium nitrite or the like. The reaction maybegin upon combination of the nitrite mud medium and the acidified mudmedium.

One embodiment of an apparatus and method in accordance with the presentinvention may provide a therapeutically effective amount of nitric oxidefrom a powder medium. The reaction may begin upon combination of thenitrite powder medium and the acidified powder medium being placed in awater bath. A water bath may provide a therapeutically effective dose ofnitric oxide over limited time, from about initial application to thirtyminutes. Reactants may include potassium nitrite, sodium nitrite or thelike.

An apparatus and method in accordance with the invention may be used fora variety of purposes, including without limitation, disinfecting andcleaning surfaces, increasing localized circulation or vasodilation,increasing systemic circulation or vasodilation, facilitating healingand growth, dispersing biofilms, reducing inflammation, and providinganalgesic benefits.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing features of the present invention will become more fullyapparent from the following description and appended claims, taken inconjunction with the accompanying drawings and experimental data.Understanding that these drawings and data depict only typicalembodiments of the invention and are, therefore, not to be consideredlimiting of its scope, the invention will be described with additionalspecificity and detail through use of the accompanying drawings and datain which:

FIG. 1 is a table illustrating the design of a subject study, includingplacement, descriptor, and amounts of relevant topical mixtures;

FIG. 2 is a table illustrating the percentage change in the peripheralperfusion index for the subject study in FIG. 1; and

FIG. 3 is a graph comparing the production of nitric oxide in a closed,control chamber with the absorption of nitric oxide through a subject'sskin.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

It will be readily understood that the components and steps of thepresent invention, as generally described and illustrated herein, couldbe arranged and designed in a wide variety of different configurations.Thus, the following more detailed description of the embodiments of thesystem and method of the present invention, as may also be representedin the drawings, is not intended to limit the scope of the invention, asclaimed, but is merely representative of various embodiments of theinvention. The illustrated embodiments of the invention will be bestunderstood by reference to the drawings, wherein like parts aredesignated by like numerals throughout.

The blood vessel of a mammal is generally lined with endothelial cells.These endothelial cells line the interior of the blood vessel. The bloodvessel may have smooth muscle around it. The endothelial cells liningthe interior of a blood vessel may include the enzyme endothelial nitricoxide synthase. The amino acid Arginine (the L-form of Arginine) may beabsorbed from the gastrointestinal tract and go into the endothelialcells. The L-Arginine is then converted to nitric oxide by theendothelial nitric oxide synthase. The nitric oxide causes the bloodvessel to open up, or vasodilation, resulting from relaxation of thesmooth muscle cells within the blood vessel walls (particularly in largearteries, arterioles, and large veins).

Some nitric oxide also enters the blood stream and binds to the redblood cells, or hemoglobin. The binding of nitric oxide to hemoglobinresults in methemoglobin.

Methemoglobin has a reduced ability to release oxygen to body tissues.Some methemoglobin in the blood stream is normal and not dangerous(generally less than I % of hemoglobin). Too much methemoglobin canresult in methemoglobinemia and even death.

The body is able to metabolize methemoglobin. The nitric oxide may beremoved from the hemoglobin and kept in the red blood cell for lateruse. Nitric oxide has a short half-life of a few seconds in the bloodunless it is bound within a red blood cell. When a blood vesselconstricts, methemoglobin may release its nitric oxide to promotevasodilation.

Nitric oxide may cause a number or tissue responses. Nitric oxide canreduce vascular smooth muscle tone. Nitric oxide can inhibit smoothmuscle proliferation and migration. Nitric oxide can promote endothelialsurvival and repair. Nitric oxide can inhibit platelet adhesion andaggregation. Nitric oxide can inhibit inflammatory mediators.

One method for measuring vasodilation is to monitor the perfusion index,or blood flow. Perfusion index is a relative assessment of the bloodpulse strength at the monitoring site in pulse oximetry. If theperfusion index trends up, it indicates vasodilation and improvedperipheral perfusion, or tissue blood flow. If the perfusion indextrends down, it indicates a decrease in peripheral perfusion. Theperfusion index (Pl) may be expressed as a rate of inflow pulsatilearterial light absorbed (AC) divided by the non-pulsatile absorbedlight, venous and non-pulsatile blood or tissue (DC). PI=AC/DC.

A person may obtain the benefits of nitric oxide therapy by utilizing atopical application that generates nitric oxide. The nitric oxide mayaffect the surface to which the topical application is applied, and maybe absorbed by a surface such as skin and affect subcutaneous tissues.

Moreover, the topical applications that may be applied can be greatlyvaried. The concentrations of the respective reactants, nitrites andreducing agents, can be varied and adjusted according to the intended ortargeted amount of nitric oxide to be produced. The viscosity of therespective, separate, initial media can vary. Generally, applicationswith thinner viscosities will produce a burst of nitric oxide, but not avery prolonged production of nitric oxide. Increasing the viscosity ofthe separate media, and the resultant topical application, can prolongand slow the production of nitric oxide, even to the point of virtuallystopping the reaction.

The viscosity of topical mixtures, and the respective, separate mediums,can vary from serums, to gels, to thixotropic substances, to any degreeof viscosity desired or intended for a specific use.

The production of nitric oxide may also be prolonged by inhibiting thenitric oxide reaction on a chemical level. For example, a salt buffermay be added to one of the separate, initial media. The salt buffer maybe any suitable salt, such as sodium chloride, calcium chloride, or thelike. In appropriate amounts, the salt buffer acts as a competitor withthe nitric oxide reactants. This can result in prolonging the productionof nitric oxide without increasing the viscosity of the separate,initial media, or the resultant topical application mixture.

In one embodiment, two individual, separate, component media areprovided. The first medium is a nitrite medium and generally providesthe nitrite reactants in a suitable form, such as sodium nitrite,potassium nitrite, or the like. The second medium is an acidified mediumand generally provides at least one acidic reactant in a suitable form,such as citric acid, lactic acid, ascorbic acid, phytic acid, aceticacid, or the like. Reaction rate and pH control are best achieved byusing a mixture of multiple food-grade acids. When approximately equalamounts of the two individual components (media) are combined into atopical mixture, a reaction is initiated that produces nitric oxide.

Two separate containers may be provided, each container is capable ofdispensing a suitable amount of a given medium (one of the two to bemixed). The containers may be identical in structure and composition,but need not necessarily be so. The containers may dispense the mediumby a pump action, such as is common with lotions and soaps. Thecontainers may dispense the medium by a squeezing or shaking action,such as is common with viscous or thixotropic shampoos, condiments,colloidal suspensions, gels, and other compositions.

The medium may be any suitable medium for containing and dispensing thereactants, for example, the medium may be a gel, a serum, a lotion, or afacial mud. A gel may be obtained by including a water-soluble polymer,such as methyl cellulose available as Methocel™, in a suitable solution.A serum may be obtained by including polyacrylate in a suitablesolution. A lotion used to suspend the reactants for a nitrite lotionmedium and an acidified lotion medium may be selected such as theJergens® brand hand and body lotion. A facial mud may be used to suspendthe reactants, for example, a facial mud medium and an acidified mudmedium may be combined to produce a facial mud topical mixture. For bestresults, the media holding a matched pair of reactants should beessentially the same. The chemical characteristics of the media may notbe strictly identical, but the physical compositions should beessentially the same so as to mix readily and not inhibit the reaction.

For example, a nitrite gel medium may have a slightly acidic to neutralpH while an acidified gel medium may have a more acidic pH than thecorresponding nitrite gel medium. Using a nitrite gel medium with anacidified lotion medium may not provide optimal results.

Using different media may not provide the best rates for desiredresults, but would probably not be harmful or dangerous.

Generally, a topical application of nitric oxide may be provided bymixing equal amounts of a nitrite medium and an acidified medium. Thetopical mixture is then applied to the intended surface. The mixture maybe applied to a person's healthy skin, or an open wound.

In one embodiment, a nitrite medium and an acidified medium are combinedto initiate the production of nitric oxide and the combination is placedon a user's skin. The nitric oxide contacts the surface of the skin andis absorbed transdermally. Thus, nitric oxide therapy can be appliedlocally for a wide variety of applications. Also, local application ofthe nitric oxide therapy can provide a prolonged, systemic effect.

The topical mixture provides nitric oxide to the intended surface. Asthe nitrite medium is mixed with the acidified medium, the reduction ofnitrite by the acid(s) leads to the release of nitric oxide. Theexposure to nitric oxide may serve a variety of purposes.

A topical mixture that produces nitric oxide may be used forantimicrobial, antifungal, or similar cleaning purposes. Infectiousdiseases are caused by pathogens such as bacteria, viruses, and fungi.Antibacterial soaps can kill some bacteria, but not necessarily allbacteria. A topical mixture as described has been shown to kill as manyas, and more, bacteria compared to commercially available antibacterialsoaps or hospital-based instant hand antiseptics.

The nitrite medium may be formulated in any suitable medium and theconcentration of reactants can be adjusted as desired as long as theintended reaction and sufficient concentrations of nitric oxide isobtained. For example, a suitable tank may be charged withdistilled/deionized water (94.94% w/w) at room temperature (20″-25° C.).Sodium nitrite (3.00% w/w) and Kathan CG (0.05% w/w) may be dissolved inthe water. Methocel™ (HPMC, cold dispersable; 1.75% w/w) may be stirredinto the water until no lumps are present. Sodium hydroxide (ION toapproximately pH 8; 0.09% w/w) may be rapidly stirred into the water tothicken, and care should be taken to avoid trapping air bubbles that canoccur as a result of higher shear mixing.

EDTA, Na4 salt (0.10% w/w) may be stirred into the water untildissolved. Citric acid (crystalline; 0.08% w/w) may be added to adjustthe mixture to a pH of 6.0. Small quantities of sodium hydroxide may beused to adjust the pH as needed. The individual percentages may beadjusted as desired for the best results.

In a separate embodiment, a nitrite medium may be formulated as follows:water, 94.8945%; sodium nitrite, 3.0000%; hydroxypropyl methylcellulose,1.0000%; phenoxyethanol, 0.4500%; caprylyl glycol, 0.2500%;ethylhexylglycerin, 0.1500%; hexylene glycol, 0.1500%; tetrasodium EDTA,0.1000%; butylene glycol, 0.0015%; grape (Vitis vinifera) seed extract,0.0010%; camellia sinensis leaf extract, 0.0010%; panax ginseng rootextract, 0.0010%; and Chamomilla recutita (matricaria) flower extract,0.0010%.

In a separate embodiment, a nitrite medium may be formulated as follows:water, 74%; caprylic/capric triglyceride, 10%; sodiumacrylate/acryloyldimethyltaurate/dimethylacrylamide crosspolymer,isohexadecane, polysorbate 60, 5%; cetearyl alcohol, 4%; sodium nitrite,3%; algae extract, Artemisia vulgaris extract, 3%; and phenoxyethanol,caprylyl glycol, ethylhexyglycerin, hexylene glycol, 1%.

In a separate embodiment, a nitrite medium may be formulated as follows:water, 93.1430%; sodium nitrite, 3.0000%; sodium bicarbonate, 1.7500%;hydroxypropyl methylcellulose, 1.0000%; phenoxyethanol, 0.4500%;caprylyl glycol, 0.2500%; ethylhexylglycerin, 0.1500%; hexylene glycol,0.1500%; tetrasodium EDTA, 0.1000%; butylene glycol, 0.0015%; glycerin,0.0015%; morus alba root extract, 0.0010%; serenoa serrulata fruitextract, 0.0010%; Rosmarinus officinalis (rosemary) leaf extract,0.0010%; and Urtica dioica (nettle) extract, 0.0010%.

In a separate embodiment, a nitrite medium may be formulated as follows:water, 87.1470%; sodium nitrite, 9.0000%; sodium bicarbonate, 1.7500%;hydroxypropyl methylcellulose, 1.0000%; phenoxyethanol, 0.4500%;caprylyl glycol, 0.2500%; ethylhexylglycerin, 0.1500%; hexylene glycol,0.1500%; tetrasodium EDTA, 0.1000%; butylene glycol, 0.0015%; glycerin,0.0015%; morus alba root extract, 0.0010%; serenoa serrulata fruitextract, 0.0010%; Rosmarinus officinalis (rosemary) leaf extract,0.0010%; and Urtica dioica (nettle) extract, 0.0010%. This formulationfor the nitrite medium was used for the gel labeled “C” in FIG. 1.

In a separate embodiment, a nitrite medium may include appropriatepercentages of the following: water; glycerin; glyceryl stearate,cetearyl alcohol, sodium stearoyl lactylate; sodium nitrite; santalumalbum (sandalwood) wood extract, phellodendrom amurense bark extract,hordeum distichion (barley) extract; algae extract, Artemisia vulgarisextract; cetearyl alcohol; sodiumacrylate/acryloyldimethyltaurate/dimethylacrylamide crosspolymer,isohexadecane, polysorbate 60; phenoxyethanol, caprylyl glycol,ethylhexylglycerin, hexylene glycol; cetyl esters; cyclopentasiloxane;hydrolyzed align, chlorella vulgaris extract, sea water, water;dimethicone; capric/caprylic triglycerides, linoleic acid, soy sterols,soy phospholipids; olea europaea (olive) fruit unsaponifiables; xanthangum; tocopheryl acetate; and hyaluronic acid.

In a separate embodiment, a nitrite medium may include appropriatepercentages of the following: water; glyceryl stearate, cetearylalcohol, sodium stearoyl, lactylate; sodium nitrite; cetearyl alcohol;cety esters; cyclopentasiloxane; and dimethicone.

The acidified medium may be formulated in any suitable carrier and theconcentration of the reactants can be adjusted as desired as long as theintended reaction and sufficient concentrations of nitric oxide areobtained. For example, a suitable tank may be charged withdistilled/deionized water (89.02% w/w) at room temperature (20″-25° C.).Kathan CG (0.05% w/w) may be dissolved in the water. Methocel™ (HPMC,cold dispersable; 1.75% w/w) may be stirred into the water until nolumps are present. Sodium hydroxide (ION to approximately pH 8; 0.09%w/w) may be rapidly stirred into the water to thicken, and care shouldbe taken to avoid trapping air bubbles that can occur as a result ofhigher shear mixing.

EDTA, Na4 salt (0.10% w/w) may be stirred into the water untildissolved. Stirring may continue until the Methocel™ is completelyhydrated. Lactic acid (85% liquid solution; 3.00% w/w) and ascorbic acid(USP, crystalline; 3.00% w/w) may be stirred in until completelydissolved. Citric acid (crystalline; 3.00% w/w) may be added to adjustthe mixture to a pH of 6.0. Small quantities of sodium hydroxide may beused to adjust the pH as needed. The individual percentages may beadjusted as desired for the best results.

In a separate embodiment, an acidified medium may be formulated asfollows: water, 90.4695%; lactic acid, 5.0000%; citric acid, 2.500%;hydroxypropyl methylcellulose (or polyacrylate crosspolymer-6), 1.400%;phenoxyethanol, 0.2250%; caprylyl glycol, 0.1250%; tetrasodium EDTA,0.1000%; ethylhexylglycerin, 0.0750%; hexylene glycol, 0.0750%; sodiumhydroxide, 0.0250%; butylene glycol, 0.0015%; grape (Vitis vinifera)seed extract, 0.0010%; camellia sinensis leaf extract, 0.0010%; panaxginseng root extract, 0.0010%; and Chamomilla Recutita (matricaria)flower extract, 0.0010%.

In a separate embodiment, an acidified medium may be formulated asfollows: water, 88.2695%; lactic acid, 5.0000%; citric acid, 3.5000%;glycerin, 1.5750%; hydroxypropyl methylcellulose, 1.0000%;phenoxyethanol, 0.2250%; caprylyl glycol, 0.1250%; tetrasodium EDTA,0.1000%; ethylhexylglycerin, 0.0750%; hexylene glycol, 0.0750%; sodiumhydroxide, 0.0250%; hydroxypropyl cyclodextrin, 0.0200%; palmitoyltripeptide-38, 0.0050%; butylene glycol, 0.0015%; morus alba rootextract, 0.0010%; serenoa serrulata fruit extract, 0.0010%; Rosmarinusofficinalis (rosemary) leaf extract, 0.0010%; and Uritica dioica(nettle) extract, 0.0010%.

In a separate embodiment, an acidified medium may be formulated asfollows: water, 84.7735%, lactic acid, 7.0000%; citric acid, 5.0000%;glycerin, 1.5750%; hydroxypropyl methylcellulose, 1.0000%;phenoxyethanol, 0.2250%; caprylyl glycol, 0.1250%; tetrasodium EDTA,0.1000%; ethylhexylglycerin, 0.0750%; hexylene glycol, 0.0750%; sodiumhydroxide, 0.0250%; hydroxypropyl cyclodextrin, 0.0200%; palmitoyltripeptide-38, 0.0050%; and butylene glycol, 0.0015%. This formulationfor the acidified medium was used for the gel labeled “C” in FIG. 1.

In a separate embodiment, each medium may include appropriate amounts ofthe respective reactants and appropriate percentages of the following:water; glycerin; cellulose gum; celery juice extract; algae extract,Artemisia vulgaris extract; and phenoxyethanol, caprylyl glycol,ethylhexylglycerin, hexylene glycol.

Ranges of acceptable constituents for each medium may include plus orminus 15% with respect to any constituent.

The use of at least two acids in producing the acidified medium mayimprove the shelf life of the acidified medium. Generally maintaining apH of from about 3 to about 5 or above (so long as not too caustic forskin) has been found very useful in maintaining the shelf life of theproduct.

A topical mixture that produces nitric oxide has been shown to beeffective in cleaning and disinfecting hands. For example, three sets ofvolunteers, with approximately 26 people in each set, participated in atest to determine the effectiveness of nitric oxide as a cleaning anddisinfecting agent. The right and left hands of each person in each setof volunteers were swabbed with cotton-tipped applicators prior to anytype of washing. The applicators were plated onto nutrient blood agarpetri dishes using the three corner dilution method.

Each set of volunteers washed their hands using separate soaps forwashing. The first set of volunteers washed their hands for thirty (30)seconds using a topical mixture of equal parts of nitrite gel medium andacidified gel medium as described herein above. The second set ofvolunteers washed their hands for thirty (30) seconds using a commercialanti-bacterial agent Avagard™D. The third set of volunteers washed theirhands for fifteen (15) seconds using Dial™ Complete Foaming Hand Wash,and then rinsed for fifteen (15) seconds and dried.

The right and left hands of each person in each set of volunteers wereswabbed again with cotton-tipped applicators after washing. Theapplicators were plated onto nutrient blood agar petri dishes using thethree corner dilution method. All the blood agar petri dishes wereincubated for forty-eight (48) hours at 35° C. The results weretabulated based on a grading scale of bacteria colonization. The testingshowed that a topical mixture that produces nitric oxide reduced therelative bacterial content by approximately 62%. Avagard™D reduced therelative bacterial content by approximately 75%. Dial™ Complete FoamingHand Wash reduced the relative bacterial content by approximately 33%.Thus, a topical mixture that produces nitric oxide was found to beapproximately twice as effective at cleaning and disinfecting hands thanDial™ Complete Foaming Hand Wash and almost as effective as Avagard™D.

It has been determined that the dose required to kill bacteria on asurface, such as a person's skin, is at least approximately 320 ppm ofnitric oxide. A topical gel mixture of approximately three (3) grams ofnitrite gel medium and approximately three (3) grams of acidified gelmedium that produces nitric oxide has been shown to deliverapproximately 840 ppm of nitric oxide. Similarly, a topical gel mixtureof approximately three (3) grams of nitrite lotion medium andapproximately three (3) grams of acidified lotion medium that producesnitric oxide has been shown to deliver approximately 450 ppm of nitricoxide.

A topical mixture that produces an effective amount of nitric oxide hasbeen shown to be effective in promoting vasodilation in generallyhealthy individuals, including systemic vasodilation. Varying amounts ofa topical mixture of gel mediums were applied to the healthy skin ofvarious individuals. Gels used to evaluate transdermal absorption ofnitric oxide and promotion of vasodilation were formulated in varyingviscosities or thicknesses, and with varying concentrations of thenitric oxide reactants. Thus, the possible amount of nitric oxide thatcould be produced was varied between the separate topical mixtures, andthe production of nitric oxide was evaluated depending on theviscosities of the gels.

It was determined that the viscosity of the gels used to form thetopical mixture should be optimized to provide a lasting, prolongedproduction of nitric oxide and promote transdermal absorption of nitricoxide. As noted in FIG. 1, Study 8 utilized Gel A (the most viscousgel), but the production of nitric oxide was noticeably slowed unlessthe application site was kept “wet.” The process of keeping the site wetdiluted Gel A, lowering the viscosity, but also promoted production ofnitric oxide. Thus, a topical mixture that was too viscous would dry outand decrease or stop the production of nitric oxide. Also, a topicalmixture that was too thin having little viscosity would provide only ashort burst production of nitric oxide and not enough transdermalabsorption of nitric oxide.

Specifically, topical mixtures identified as A, B and C were used in theexperimentation. The nitrite gel medium and the reducing agent oracidified medium for each topical mixture contained varying amounts ofreactants. Likewise, the viscosities of the gels were varied. Theviscosity of the topical mixtures A, B and C proceed from highest tolowest, with A being the most viscous and C being the least viscous. Thesite and amount of the respective gels applied to each study, or person,is shown in FIG. 1.

Hemodynamic measurements for each study were recorded at 30 minutes and15 minutes prior to application of the respective gels. Hemodynamicmeasurements for each study were recorded again at 5, 15, 30, 45, 60,75, 90, and 120 minutes after application of the respective gels.

For example, Table I below shows the hemodynamic changes for Study 10with respect to systolic pressure, diastolic pressure, mean arterialblood pressure (MABP), and heart rate.

TABLE I Systolic Pressure Diastolic Pressure MABP HR Pre 30 mins. 130 80113 75 Pre 15 mins. 132 81 115 76 Post 5 mins. 130 78 113 78 Post 15mins. 128 76 111 72 Post 30 mins. 122 70 105 68 Post 45 mins. 115 69 10070 Post 60 mins. 112 67 97 64 Post 75 mins. 120 70 103 70 Post 90 mins.115 68 99 68 Post 120 mins. 119 70 103 69

Table 2 below shows the Peripheral Perfusion Index (P1) for Study 10.

TABLE 2 PI Pre 30 mins. 4.0 Pre 15 mins. 3.8 Post 5 mins. 4.2 Post 15mins. 6.0 Post 30 mins. 7.0 Post 45 mins. 7.8 Post 60 mins. 9.0 Post 75mins. 6.0 Post 90 mins. 8.0 Post 120 mins. 7.0

Table 3 below shows the methemoglobin (metHg) percentages for Study 10.

TABLE 3 metHg % Pre 30 mins. 0.2 Pre 15 mins. 0.2 Post 5 mins. 0.3 Post15 mins. 0.5 Post 30 mins. 0.5 Post 45 mins. 0.5 Post 60 mins. 0.5 Post75 mins. 0.5 Post 90 mins. 0.5 Post 120 mins. 0.5

To ensure that methemoglobin was not “pulling” or gathering in alocalized area of the body, methemoglobin percentage was measured fromthe middle finger, index finger, and thumb of the subject's right andleft hands, and from the middle toe, second toe, and big toe of thesubject's right and left feet prior to the application of the topicalmixture and one hour after application of the topical mixture thatproduced nitric oxide. No significant pulling was found. The results areprovided in Table 4 below.

TABLE 4 metHg % metHg % (prior) (one hour) Left hand, Middle finger 0.20.5 Left hand, Index finger 0.2 0.5 Left hand, Thumb 0.0 0.4 Right hand,Middle finger 0.0 0.5 Right hand, Index finger 0.0 0.5 Right hand, Thumb0.0 0.5 Left foot, Middle toe 0.2 0.5 Left foot, Second toe 0.2 0.4 Leftfoot, Big toe. 0.1 0.5 Right foot, Middle toe 0.0 0.4 Right foot, Secondtoe 0.0 0.4 Right foot, Big toe. 0.0 0.4

As shown in FIG. 2, the change in peripheral perfusion index across allstudies shows that a topical mixture that produces an effective amountof nitric oxide increases vasodilation systemically. Also, a topicalmixture that produces nitric oxide more quickly may have less sustainedsystemic effects. A topical mixture that produces nitric oxide over amore prolonged period of time can have sustained hemodynamic, systemiceffects for up to two hours. The methemoglobin levels were consistentthroughout the body demonstrating a whole body distribution of nitricoxide, while maintaining acceptable levels of methemoglobin.

The kinetics for the formation of nitric oxide were also evaluated todetermine if nitric oxide was absorbed better through some skin areas ascompared to other. For example, Table 5 below shows the nitric oxideproduction comparison between a “control” amount of gel A (production ofnitric oxide in a closed container) and the production of nitric oxideusing the same amount of gel A applied to the chest of Study 4.

TABLE 5 Control-NO (ppm) Chest-NO (ppm)  0 mins. 740 0  7 mins. 756 0 13mins. 500 156 23 mins. 352 58 34 mins. 104 26 45 mins. 21 2

As another example, Table 6 below shows the nitric oxide productioncomparison between a “control” amount of gel A (production of nitricoxide in a closed container) and the production of nitric oxide usingthe same amount of gel A applied to the right leg of Study 5.

TABLE 6 Control-NO (ppm) Chest-NO (ppm)  0 mins. 877 34  8 mins. 150 1017 mins. 101 6 30 mins. 82 4 37 mins. 40 5 40 mins. 37 5 55 mins. 12 465 mins. 17 1 97 mins. 10 0

As shown in FIG. 3, and as substantiated by the data in Tables 5 and 6,nitric oxide produced from application of a topical mixture is absorbedthrough the skin of a person. As established from the other data fromFIG. 2 and Tables 1-4, the nitric oxide absorbed through the skinpromotes systemic vasodilation without substantial risk ofmethemoglobinemia.

The topical mixture applied to a given site would dry out approximately30 minutes after application, but the hemodynamic effects could still bemeasured for approximately an additional 60 to 90 minutes.

The skin may be covered with a barrier, cover, or bandage afterapplication of the nitric oxide producing topical mixture. Such abarrier can serve many purposes, including, maintaining the moisturecontent in the topical mixture, preventing debris or contaminants fromgetting into the topical mixture or on the skin, and promoting thetransdermal absorption of the nitric oxide produced.

The amount of nitric oxide that is produced and absorbed transdermallymay vary based on site applications, and certainly varies depending onthe amount of nitric oxide producing reactants present in the topicalmixture applied and the duration of the nitric oxide producing reaction.Generally, the amount of nitric oxide absorbed transdermally betweeninitial application of the topical mixture and approximately 60-90minutes is between approximately 1000 ppm and 2000 ppm, and can bebetween 500 ppm and 2500 ppm.

One possible application for this data is in the treatment of peripheralartery disease (PAD). Peripheral artery disease is a common disorderwith a progressive reduction in lower limb blood flow, which in severecases leads to chronic pain, gangrene, and eventually limb loss.Peripheral artery disease is characterized by progressive occlusion oflarge and medium-sized arteries outside the cardio-pulmonary andcerebral vascular systems. Dysregulation of nitric oxide production inthe endothelial lining of blood vessels is felt to be a majorcontributor.

Application of topical mixtures that produce nitric oxide may become aless expensive and faster mechanism to improve bioavailability ofsystemic nitric oxide pools as compared to currently availabletreatments.

Likewise, application of topical mixtures that produce nitric oxide maybecome a therapeutic possibility in the treatment of systemichemodynamic disorders of all kinds.

A mixture that produces nitric oxide may be used for a variety ofpurposes and ailments, including without limitation, diabetic andnon-healing wounds or lesions, leprosy, tuberculosis, malaria,anti-parasitic uses, etc. In one embodiment, the mixture can be used asa topical application on healthy skin. There are combinations of usesalso possible.

A topical mixture that produces nitric oxide may be used to help healvarious kinds of wounds. Tests have been performed wherein a topicalmixture that produces nitric oxide as described herein is appliedregularly to an open wound that is generally resistant to healing. Thewound was seen to show significant healing within a few weeks.

For example, a person in Canada had poor circulation and unresponsivediabetic ulcers on the person's feet. The person was immobilized and ina wheel chair, and had been scheduled for amputation to remove theperson's foot about a month after this experiment began. A topicalmixture that produces nitric oxide was applied to the diabetic ulcersonce a day. The person soaked the effected foot in a footbath solutionthat produces nitric oxide for approximately twenty minutes once everyfour days. Within two weeks the person was able to walk and go out inpublic. Within 4-6 weeks, the person was mobile and had achieved asubstantially complete recovery. Meanwhile, the scheduled amputation wascancelled.

The nitric oxide may promote the healing of diabetic ulcers in at leasttwo ways: the surface of the wound area is cleaned and sterilized, andthe underlying vascularization of the area is improved, vasodilation.The nitric oxide may also help disperse any biofilms created as a resultof the diabetic ulcer. Thus, healing of the wound is promoted from thesurface and below the surface.

A topical mixture that produces nitric oxide may be used for localizedanalgesic purposes. The analgesic effect nitric oxide may be providedvia topical application.

A topical mixture that produces nitric oxide may be used for treatingerectile dysfunction. Nitric oxide acts as a vasodilator. The nitricoxide provided by the topical mixture and absorbed transdermally maystimulate and increase blood flow to the treated area. Increased bloodflow may also help performance of muscles or muscle groups that receivenitric oxide transdermally.

A topical mixture that produces nitric oxide may be used foranti-inflammatory purposes.

A topical mixture that produces nitric oxide may also be used todisperse a biofilm. Acne is an example of a biofilm. Biofilms arecolonies of dissimilar organisms that seem to join symbiotically toresist attack from antibiotics. Nitric oxide signals a biofilm todisperse so antibiotics can penetrate the biofilm. It is also believedthat nitric oxide interferes with the uptake of iron.

It has been shown that a topical mixture that produces nitric oxide willalso treat acne. Acne is a biofilm. As an example, at least two peoplehad a considerable amount of acne on their faces. A topical mixture thatproduces nitric oxide was applied to the acne twice a day. Within twodays a marked amount of the acne had been cleared and inflammation wasgreatly reduced. Within two weeks the acne was substantially gone. Atopical mixture that produces nitric oxide will treat the surface of theacne, as well as treating the underlying causes of the acne. Theresultant nitric oxide will disperse the biofilm, act as an antisepticmicrobicide agent against bacteria, and promote vascularizationsubcutaneously to promote cleansing and healing.

Other types of biofilms can include methicillin-resistant Staphylococcusaureus (MRSA) and gangrene. For example, two people had unresponsive,non-healing wounds on their lower legs, including biofilms that furtherprevented healing. A topical mixture that produces nitric oxide wasapplied to the wounds twice a day. The person soaked and washed theeffected wound in a bath solution that produces nitric oxide forapproximately twenty minutes once every three to four days. Withinapproximately four weeks the wound had closed and the person hadachieved a substantially complete recovery.

A topical mixture that produces nitric oxide can treat the surface ofthe skin and help promote healing, but demonstrating how to effectivelypromote vasodilation via transdermal absorption of nitric oxide showsthat the underlying, subcutaneous issues preventing healing can also betreated. The nitric oxide absorbed transdermally can act subcutaneouslyto disperse a biofilm, act as an antiseptic microbicide agent againstbacteria, reduce or eliminate inflammation, and promote vascularizationsubcutaneously to promote cleansing and healing.

It was shown that a topical mixture that produces nitric oxide will killsquamous cells, pre-cancerous cells, if the concentration of nitricoxide is high enough. Tests intending to show that a topical mixturethat produces nitric oxide would grow hair based in part on the increaseof blood flow that accompanies application of nitric oxide actuallyshowed that nitric oxide in as high doses provided as described hereinabove did kill squamous cells.

A topical mixture that produces nitric oxide may be used for morecosmetic purposes. The nitric oxide dose provided via topicalapplication may reduce the appearance of wrinkles.

A variety of topical mixtures may be obtained by combining one ofvarious types of nitrite mediums with one of various types of acidifiedmediums. For example and without limitation, a nitrite gel medium may becombined with an acidified serum medium, or vice versa. Likewise, othercombinations of mediums are possible.

It may also be noticed that using nitric oxide producing gels as atechnique to wash and sterilize hands over a significant number of daysmay lead to vasodilation of the blood vessels in the hands.

The present invention may be embodied in other specific forms withoutdeparting from its spirit or essential characteristics. The describedembodiments are to be considered in all respects only as illustrative,and not restrictive. The scope of the invention is, therefore, indicatedby the appended claims, rather than by the foregoing description. Allchanges which come within the meaning and range of equivalency of theclaims are to be embraced within their scope.

What is claimed and desired to be secured by United States LettersPatent is: 1-19. (canceled)
 20. A composition for increasing systemicvasodilation in a mammal with an epidermal surface and a bloodstreamhaving an initial systemic vasodilation of the bloodstream, comprising:a first medium comprising a nitrite compound; a second medium comprisingat least two reducing agents; wherein the first medium and the secondmedium are adapted to be mixed into a mixture by a user during use,thereby initiating production of nitric oxide in an effective amountsuch that application of the mixture onto a first location of anepidermal surface of a mammal is configured to deliver at least 500 ppmof nitric oxide transdermally within 10 minutes after the application,thereby increasing the systemic vasodilation of the bloodstream at asecond, untreated location of the mammal as compared to the initialsystemic vasodilation during use.
 21. The composition of claim 20,wherein the epidermal surface is healthy skin.
 22. The composition ofclaim 20, wherein a mixture of the first medium and the second medium,in use, includes a salt buffer concentration for prolonging theproduction of nitric oxide, wherein the salt used in the salt bufferincludes at least one salt selected from a group consisting of sodiumchloride and calcium chloride.
 23. The composition of claim 20, whereinthe mixture is adapted for treating the mammal for a symptom ofperipheral artery disease.
 24. The composition of claim 20, wherein thefirst medium includes at least sodium nitrite.
 25. The composition ofclaim 20, wherein the first medium includes at least potassium nitrite.26. The composition of claim 20, wherein the second medium includes atleast two reducing agents selected from a group consisting of citricacid, ascorbic acid, phytic acid, acetic acid, formic acid, and lacticacid.
 27. The composition of claim 20, wherein the first and secondmediums further include a thickening agent.
 28. The method of claim 20,wherein the first medium further comprises sodium bicarbonate.
 29. Acomposition for treating a systemic disorder, comprising: a first mediumcomprising a nitrite compound; and a second medium comprising a reducingagent; wherein the first medium and the second medium are adapted to bemixed into a mixture by a user during use thereby initiating productionof nitric oxide in an effective amount such that application of themixture onto a first location of a treatment surface of a subject isconfigured to increase systemic vasodilation of the bloodstream at asecond, unapplied location of the subject to treat the systemic disorderduring use.
 30. The composition of claim 29, wherein application of themixture is adapted to treat symptoms of the systemic disorder peripheralartery disease.
 31. The composition of claim 29, wherein the secondmedium includes at least two reducing agents selected from a groupconsisting of citric acid, ascorbic acid, phytic acid, acetic acid,formic acid, and lactic acid.
 32. The composition of claim 29, whereinthe mixture includes sodium chloride.
 33. The composition of claim 29,wherein the mixture includes calcium chloride.
 34. A composition forincreasing vasodilation in a subject comprising: a first mediumcomprising a nitrite compound; a second medium comprising a reducingagent; wherein the first medium and the second medium are adapted to bemixed into a mixture by a user during use, wherein the first medium andsecond medium are configured such that the mixture includes a salt forprolonging the production of nitric oxide, wherein during use themixture is configured to initiate production of nitric oxide such thatapplication of the mixture onto a first location of a treatment surfaceof a subject is configured to deliver an effective amount of nitricoxide transdermally, and adapted to increase the systemic vasodilationof the bloodstream at a second, untreated location of the subject ascompared to the initial systemic vasodilation during use.
 35. Thecomposition of claim 34, wherein the second medium comprises citricacid.
 36. The composition of claim 34, wherein the second mediumcomprises ascorbic acid.
 37. The composition of claim 34, wherein thesecond medium comprises phytic acid.
 38. The composition of claim 34,wherein the second medium comprises acetic acid.
 39. The composition ofclaim 34, wherein the second medium comprises formic.
 40. Thecomposition of claim 34, wherein the second medium comprises lacticacid.